miRNA在鸡胚胎干细胞与鸡胚成纤维细胞中表达差异的研究  

Differential expression of micro RNAs in chicken embryonic stem cells and chicken embryo fibroblast cells

作　　者：冯娜[1] 陈胜锋[1] 王丙云[1] 陈志胜[1] 计慧琴[1] 

机构地区：[1]佛山科学技术学院动物医学系,广东佛山528231

出　　处：《中国兽医科学》2016年第12期1596-1601,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31201780);广东省自然科学基金项目(2015A030313632)

摘　　要：为筛选在鸡胚胎干细胞(c ESCs)和鸡胚成纤维细胞(CEFs)中差异表达的micro RNA(miRNA),并探讨miRNA在维持c ESCs多能性过程中的作用,分别体外培养c ESCs、CEFs,应用miRNA芯片检测c ESCs、CEFs的miRNA表达谱,筛选差异表达的miRNA,并采用实时荧光定量PCR进行验证,同时利用生物信息学软件对靶基因进行预测。结果显示,从已知的993条鸡miRNA中,筛选出gga-miR-1777、gga-miR-302d、gga-miR-1607、gga-miR-1599和gga-miR-1576等19条在c ESCs中高水平表达的miRNA。实时荧光定量PCR检测结果显示,gga-miR-1777、gga-miR-302d的表达与miRNA芯片结果相符。生物信息学分析显示,gga-miR-1777、gga-miR-302d的靶基因中包含细胞分化相关的基因。研究结果提示,c ESCs中高水平表达的miRNA可能通过抑制分化基因的表达来维持c ESCs的多能性。To screen significant mi RNAs which differentially expressed in chicken embryonic stem cells（c ESCs）and chicken embryo fibroblast cells（CEFs）,and to evaluate its roles involved in pluripotency in c ESCs,the mi RNA expression profiles of c ESCs and CEFs were detected by mi RNA microarray.The differentially expressed mi RNAs were identified based on microarray dates and verified by quantitative real-time PCR.Meanwhile,the potential target genes of identified mi RNAs were predicted by bioinformatics software.several mi RNAs,such as gga-mi R-1777,gga-mi R-302 d,gga-mi R-1607,gga-mi R-1599 and gga-mi R-1576,etc,which had high expression in c ESCs,were screened out from 993 known mi RNAs in chicken.The expression of gga-mi R-1777 and gga-mi R-302 d detected by quantitative real-time PCR as consistent with mi RNA microarray dates.The analysis results by bioinformatics software showed that some of potential target genes of gga-mi R-1777,and gga-mi R-302 d were involved in cell differentiation.The results indicated that high expression of mi RNAs may be maintained pluripotency by suppressing genes related to differentiation in c ESCs.

关 键 词：MICRORNAS 鸡胚胎干细胞 MIRNA芯片 多能性 

分 类 号：S852.163[农业科学—基础兽医学]